CLP model was induced as we previously described [4 (link)]. Briefly, animals were anesthetized with 2% sodium pentobarbital (50 mg/kg; Sigma Chemical Co, St. Louis, MO, USA) by intraperitoneal (i.p.) injection and a 1-cm ventral midline laparotomy was performed. After then, the cecum was carefully exposed and ligated with a 4.0 silk suture, about 0.5 cm below the ileocecal valve. Subsequently, the cecum was perforated with 22–gauge needle and gently compressed to extrude a small amount of feces. Finally, the cecum was returned to the peritoneal cavity and the laparotomy was closed with 4.0 silk sutures. Immediately after the operation, animals received fluid resuscitation with normal saline solution (subcutaneously, 20 ml/kg of body weight) and antibiotic therapy (ertapenem, 20 mg kg-1; Merck Research Laboratory, USA). All mice were returned to their cages with free access to food and water. For sham group, animals were treated identically without ligation or puncture of the cecum.
Modeling Sepsis in Rodents: LPS and CLP
CLP model was induced as we previously described [4 (link)]. Briefly, animals were anesthetized with 2% sodium pentobarbital (50 mg/kg; Sigma Chemical Co, St. Louis, MO, USA) by intraperitoneal (i.p.) injection and a 1-cm ventral midline laparotomy was performed. After then, the cecum was carefully exposed and ligated with a 4.0 silk suture, about 0.5 cm below the ileocecal valve. Subsequently, the cecum was perforated with 22–gauge needle and gently compressed to extrude a small amount of feces. Finally, the cecum was returned to the peritoneal cavity and the laparotomy was closed with 4.0 silk sutures. Immediately after the operation, animals received fluid resuscitation with normal saline solution (subcutaneously, 20 ml/kg of body weight) and antibiotic therapy (ertapenem, 20 mg kg-1; Merck Research Laboratory, USA). All mice were returned to their cages with free access to food and water. For sham group, animals were treated identically without ligation or puncture of the cecum.
Corresponding Organization : First Affiliated Hospital of Zhengzhou University
Variable analysis
- LPS injection (Escherichia coli endotoxin 0111: B4, Lot # 064M4125V, Sigma, Shanghai, China, 5 mg/kg)
- CLP model (cecal ligation and puncture)
- Unstated
- Experienced investigator to keep the model stable
- Anesthesia with 2% sodium pentobarbital (50 mg/kg; Sigma Chemical Co, St. Louis, MO, USA) by intraperitoneal (i.p.) injection
- Fluid resuscitation with normal saline solution (subcutaneously, 20 ml/kg of body weight)
- Antibiotic therapy (ertapenem, 20 mg kg^-1; Merck Research Laboratory, USA)
- Access to food and water
- CLP model as previously described [4 (link)]
- Sham group, where animals were treated identically without ligation or puncture of the cecum
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